The free amino acid profile of 18 samples of tronchuda cabbage (Brassica oleracea L. var. costata DC) leaves, harvested at three different months, was determined by HPLC/UV−vis. The tronchuda cabbage leaves total free amino acid content varied from 3.3 to 14.4 g/kg fresh weight. Generally, arginine was the major compound, followed by proline, threonine, glutamine, cysteine, and glutamic acid. This study indicates that free amino acids are not similarly distributed: in external leaves, proline and arginine were the major free amino acids, while in internal ones, arginine was the main free amino acid, followed by threonine, glutamine, and cysteine. Significant differences were observed for valine, proline, arginine, leucine, cysteine, lysine, histidine, and tyrosine contents. The levels of some free amino acids were significantly affected by the collection period. In external leaves, this occurred with glutamic acid, serine, valine, leucine, cysteine, and ornithine contents, while in internal leaves, it occurred with aspartic acid, arginine, and total contents.

The ability of the aqueous extract of tronchuda cabbage (Brassica oleracea L. var. costata DC) external leaves to act as a
scavenger of DPPH- and reactive oxygen species (superoxide radical, hydroxyl radical and hypochlorous acid) was investigated.
A phytochemical study was also undertaken, and thirteen phenolic compounds and five organic acids were identified and quantified.
Tronchuda cabbage extracts exhibited antioxidant capacity in a concentration-dependent manner in all assays, although some
pro-oxidant effect was also noticed. The samples with higher phenolic and organic acid contents displayed the major antioxidant
potentials.

Tronchuda cabbage (Brassica oleracea L. var. costata DC) seeds were studied for their chemical composition and antioxidant capacity.
Thirteen phenolic compounds were characterized and quantified by reversed-phase HPLC-DAD-MS/MS-ESI and HPLC-DAD,
respectively: two sinapoylgentiobiose isomers, three sinapoylglucose isomers, kaempferol-3-(sinapoyl)sophorotrioside-7-glucoside, sinapoylcholine,
kaempferol-3,7-diglucoside-40-(sinapoyl)glucoside, three disinapoylgentiobiose isomers, 1,2,20-trisinapoylgentiobiose and
1,2-disinapoylglucose. Seven organic acids (aconitic, citric, ascorbic, malic, quinic, shikimic and fumaric acids) were also identified
and quantified by HPLC-UV. The aqueous extract of tronchuda cabbage seeds was investigated for its capacity to act as a scavenger
of DPPH- radical and reactive oxygen species (superoxide radical, hydroxyl radical and hypochlorous acid), exhibiting antioxidant
capacity in a concentration-dependent manner against all radicals. These results may be attributed to the high content of hydroxycinnamic
derivatives and ascorbic acid.

The changes in antioxidant compounds of Brassica oleracea L. var. costata DC seeds were monitored during
the first twelve days of seedling development. Sprouts were screened at time intervals of two days for phenolic compounds
and organic acids. The identified phenolic compounds included esters of sinapic acid with glucose, gentiobiose
and kaempferol, as well as sinapoylcholine. The organic acids were oxalic, aconitic, citric, pyruvic, malic, shikimic, and
fumaric acids. During germination, a depletion of phenolic compounds was observed, although no qualitative changes
were seen. Among individual compounds, kaempferol, choline and glucose esters of sinapic acid showed a marked decrease
between days two and six, whereas the changes in gentiobiose esters of sinapic acid were smaller. The total organic
acids content increased rapidly during the first four days, with less significant variations thereafter. Malic acid, the major
organic acid found in sprouts, greatly contributed to this result though oxalic, pyruvic, and fumaric acids also increased in
the same manner. In contrast, aconitic, citric and shikimic acids showed decreases between days two and twelve of germination.

A field experiment was carried out to investigate the effect of fertilization level on the phenolic composition of tronchuda cabbage (Brassica oleracea L. var. costata DC) external and internal leaves.
Eight different plots were constituted: a control without fertilization, one with organic matter, and six
experiments with conventional fertilizers (nitrogen, boron, and sulfur, two levels each). The phenolic
compounds were analyzed by reversed-phase HPLC-DAD. External and internal leaves revealed
distinct qualitative composition. In the internal leaves were found 15 phenolics (5 kaempferol and 10
cinnamic acid derivatives), whereas the external leaves presented 3-p-coumaroylquinic acid and 13
kaempferol derivatives. Principal component analysis (PCA) was applied to assess the relationships
between phenolic compounds, agronomical practices, and harvesting time. Samples obtained with
conventional practices were quite effectively separated from organic samples, for both types of leaves.
In general, samples developed without any fertilization presented the highest phenolics amounts:
external and internal leaves contained 1.4- and 4.6-fold more phenolic compounds than the ones
that received conventional fertilizer, respectively...

The phenolic compounds produced by in vitro shoots of Brassica oleracea L. var. costata DC were screened by
HPLC-DAD-MS/MS. Thirty seven compounds were characterized, which included chlorogenic acids, flavonoids (the majority
of them were hydroxycinnamic acid esters of kaempferol and quercetin glycosides) and hydroxycinnamic acyl glycosides
(with predominance of synapoyl gentiobiosides). The antioxidant capacity of the shoots was assessed against
DPPH radical and two reactive oxygen species (superoxide radical and hypochlorous acid). A strong concentrationdependent
antioxidative capacity was verified in the DPPH and superoxide radicals assays, but a reduced effect was noticed
against hypochlorous acid. The results obtained indicate that the in vitro production of B. oleracea var. costata
shoots can become important in the obtention of a noticeable dietary source of compounds with health protective potential.

Phenolic and organic acid profiles of aqueous extracts from Pieris brassicae material and the host kale
(Brassica oleracea L. var. acephala) leaves were determined by HPLC/UV–DAD/MSn-ESI and HPLC–UV,
respectively. The identified phenolics included acylated and nonacylated flavonoid glycosides, hydroxycinnamic
acyl gentiobiosides, and sulphate phenolics. Kale exhibited the highest content (11 g/kg lyophilized
extract), while no phenolics were identified in the butterflies or exuviae. Nine different organic
acids were characterized in the materials, with kale showing the highest amount (112 g/kg lyophilized
extract). With the exception of the exuviae extract, the rest were screened for bioactivity. Using spectrophotometric
microassays, all exhibited antiradical capacity against DPPH and NO in a concentrationdependent
way, whereas only kale and excrement extracts were active against superoxide. All displayed
activity on intestinal smooth muscle, albeit with distinct relaxation–contraction profiles. Larvae and butterfly
extracts were more efficacious for intestinal relaxation than was kale extract, whereas excrement
extract evoked only contractions, thus evidencing their different compositions. Collectively, these results
show that P. brassicae sequesters and metabolizes kale’s phenolic compounds. Moreover...

In this work, the volatile composition of kale (Brassica oleracea L. var. acephala) and its variation
during germination were monitored during the first 9 days of seedling development by headspace
solid-phase microextraction (HS-SPME) combined with gas chromatography/ion trap-mass spectrometry
(GC/IT-MS). Differences were found among the materials in the distinct analyzed periods.
A total of 66 volatile compounds, distributed in several chemical classes, were determined: alcohols,
carbonyl compounds (ketones, aldehydes, and esters), norisoprenoids, and terpenes, among
others, sulfur compounds being the most abundant group in seeds and sprouts that exhibited allyl
isothiocyanate as the major compound. Leaves of fully developed ground plant had the highest
content of norisoprenoids, alcohols, and carbonyl compounds; in opposition, they showed lower
levels of sulfur compounds, suggesting that these are important molecules for the development of
kale, whereas the others are produced mainly during its growth.

10 pages, 6 figures, 4 tables.-- Printed version published 2006.; The ability of the aqueous extract of tronchuda cabbage (Brassica oleracea L. var. costata DC) external leaves to act as a scavenger of DPPH and reactive oxygen species (superoxide radical, hydroxyl radical and hypochlorous acid) was investigated. A phytochemical study was also undertaken, and thirteen phenolic compounds and five organic acids were identified and quantified. Tronchuda cabbage extracts exhibited antioxidant capacity in a concentration-dependent manner in all assays, although some pro-oxidant effect was also noticed. The samples with higher phenolic and organic acid contents displayed the major antioxidant potentials.; The authors are grateful to Fundaçäo para a Ciência e Tecnologia (POCI/AGR/57399/2004) for financial support of this work. Vendula Vrchovská is grateful to the European Union Erasmus/Socrates II Programme for a grant (MSM 0021620822).; Peer reviewed

7 pages, 3 tables, 4 figures.; Glycosylated kaempferol derivatives from the external leaves of tronchuda cabbage (Brassica oleracea L. var. costata DC) characterized by reversed-phase HPLC-DAD-MS/MS-ESI were kaempferol 3-O-sophorotrioside-7-O-glucoside, kaempferol 3-O- (methoxycaffeoyl/caffeoyl)sophoroside-7-O-glucoside, kaempferol 3-O-sophoroside-7-O-glucoside, kaempferol 3-O-sophorotrioside-7-O-sophoroside, kaempferol 3-O-sophoroside-7-O-sophoroside, kaempferol 3-O-tetraglucoside-7-O-sophoroside, kaempferol 3-O-(sinapoyl/caffeoyl)sophoroside-7-O-glucoside, kaempferol 3-O-(feruloyl/caffeoyl)sophoroside-7-O-glucoside, kaempferol 3-O-sophorotrioside, kaempferol 3-O-(sinapoyl)sophoroside, kaempferol 3-O-(feruloyl)sophorotrioside, kaempferol 3-O-(feruloyl)sophoroside, kaempferol 3-O-sophoroside, and kaempferol 3-O-glucoside. These acylated derivatives are reported for the first time in nature, with the exception of kaempferol 3-O-(sinapoyl)sophoroside. Quantification of the identified compounds was achieved by HPLC-DAD and carried out in samples cultivated under conventional or organic practices and collected at different times. In general, samples from organic production exhibited higher total phenolics content than those from conventional practices collected in the same period.; We are grateful to Fundaçäo para a Ciência e Tecnologia (POCTI/AGR/57399/2004) for
financial support of this work.; Peer reviewed

9 pages, 5 figures, 2 tables.; A field experiment was carried out to investigate the effect of fertilization level on the phenolic composition of tronchuda cabbage (Brassica oleracea L. var. costata DC) external and internal leaves. Eight different plots were constituted: a control without fertilization, one with organic matter, and six experiments with conventional fertilizers (nitrogen, boron, and sulfur, two levels each). The phenolic compounds were analyzed by reversed-phase HPLC-DAD. External and internal leaves revealed distinct qualitative composition. In the internal leaves were found 15 phenolics (5 kaempferol and 10 cinnamic acid derivatives), whereas the external leaves presented 3-p-coumaroylquinic acid and 13 kaempferol derivatives. Principal component analysis (PCA) was applied to assess the relationships between phenolic compounds, agronomical practices, and harvesting time. Samples obtained with conventional practices were quite effectively separated from organic samples, for both types of leaves. In general, samples developed without any fertilization presented the highest phenolics amounts: external and internal leaves contained 1.4- and 4.6-fold more phenolic compounds than the ones that received conventional fertilizer...

7 pages, 6 figures, 4 tables.; The ability of tronchuda cabbage (Brassica oleracea L. var. costata DC) to act as a scavenger of the reactive nitrogen species nitric oxide and peroxynitrite was investigated. The aqueous extracts obtained from tronchuda cabbage seeds and from its external and internal leaves exhibited a concentration dependent scavenging capacity. The antioxidant potential observed against the two reactive species was as follows: seeds > external leaves > internal leaves. In order to establish a possible correlation with the chemical composition of the extracts, the activity of ascorbic and sinapic acids and kaempferol 3-O-rutinoside was also studied. Among the compounds tested, sinapic acid showed the strongest antioxidant activity against both species.; We are grateful to Fundaçäo para a Ciência e a Tecnologia (POCI/AGR/57399/2004) for
financial support of this work.; Peer reviewed

10 pages, 6 figures, 3 tables.; Tronchuda cabbage (Brassica oleracea L. var. costata DC) seeds were studied for their chemical composition and antioxidant capacity. Thirteen phenolic compounds were characterized and quantified by reversed-phase HPLC-DAD-MS/MS-ESI and HPLC-DAD, respectively: two sinapoylgentiobiose isomers, three sinapoylglucose isomers, kaempferol-3-(sinapoyl)sophorotrioside-7-glucoside, sinapoylcholine, kaempferol-3,7-diglucoside-4′-(sinapoyl)glucoside, three disinapoylgentiobiose isomers, 1,2,2′-trisinapoylgentiobiose and 1,2-disinapoylglucose. Seven organic acids (aconitic, citric, ascorbic, malic, quinic, shikimic and fumaric acids) were also identified and quantified by HPLC-UV. The aqueous extract of tronchuda cabbage seeds was investigated for its capacity to act as a scavenger of DPPH radical and reactive oxygen species (superoxide radical, hydroxyl radical and hypochlorous acid), exhibiting antioxidant capacity in a concentration-dependent manner against all radicals. These results may be attributed to the high content of hydroxycinnamic derivatives and ascorbic acid.; The authors are grateful to Fundacäo para a Ciência e Tecnologia (POCI/AGR/57399/2004) for financial support of this work.; Peer reviewed

10 pages.; The changes in antioxidant compounds of Brassica oleracea L. var. costata DC seeds were monitored during the first twelve days of seedling development. Sprouts were screened at time intervals of two days for phenolic compounds and organic acids. The identified phenolic compounds included esters of sinapic acid with glucose, gentiobiose and kaempferol, as well as sinapoylcholine. The organic acids were oxalic, aconitic, citric, pyruvic, malic, shikimic, and fumaric acids. During germination, a depletion of phenolic compounds was observed, although no qualitative changes were seen. Among individual compounds, kaempferol, choline and glucose esters of sinapic acid showed a marked decrease between days two and six, whereas the changes in gentiobiose esters of sinapic acid were smaller. The total organic acids content increased rapidly during the first four days, with less significant variations thereafter. Malic acid, the major organic acid found in sprouts, greatly contributed to this result though oxalic, pyruvic, and fumaric acids also increased in the same manner. In contrast, aconitic, citric and shikimic acids showed decreases between days two and twelve of germination.; Peer reviewed

5 pages, 3 figures, 2 tables.; A phytochemical study was undertaken on tronchuda cabbage (Brassica oleracea L. var. costata DC) cultivated under conventional and organic practices and collected at different times. Six organic acids (aconitic, citric, ascorbic, malic, shikimic, and fumaric acids) were identified and quantified by HPLC-UV. Qualitative and quantitative differences were noted between internal and external leaves. Analysis of the phenolics of the internal leaves was achieved by HPLC-DAD, and the phenolic profile obtained was revealed to be distinct from that of the external leaves. By this means were identified and quantified 11 compounds: 3-p-coumaroylquinic acid, kaempferol 3-O-sophoroside-7-O-glucoside, kaempferol 3-O-(caffeoyl)sophoroside-7-O-glucoside, kaempferol 3-O-(sinapoyl)sophoroside-7-O-glucoside, kaempferol 3-O-(feruloyl)sophoroside-7-O-glucoside, kaempferol 3-O-sophoroside, two isomeric forms of 1,2-disinapoylgentiobiose, 1-sinapoyl-2-feruloylgentiobiose, 1,2,2‘-trisinapoylgentiobiose, and 1,2‘-disinapoyl-2-feruloylgentiobiose. In general, internal leaves exhibited more constant chemical profiles.; We are grateful to the Fundaçäo para a Ciência e Tecnologia (POCI/AGR/57399/2004) for financial support of this work.; Peer reviewed